Effects of contaminants on blood group factors

ISSN: 2394-0026 (P)

ISSN: 2394-0034 (O)

Original Research Article

Evidentiary value and effects of

contaminants on blood group factors in
medico
medico-legal
grounds
Ashwini Narayan K1*, Manjunath MR2, Kusuma KN3
1

Assistant Professor, Department of Forensic Medicine, Mandya Institute of Medical Sciences,

Mandya, Karnataka, India
2
Associate Professor, Department of Pathology, Mandya Institute of Medical Sciences, Mandya,
Karnataka, India
3
PG Student, Department of Pathology, Mandya Institute of Medical Sciences, Mandya, Karnataka,
India
*Corresponding author email: narayanashwin9@gmail.com
How to cite this article: Ashwini Narayan K, Manjunath MR, Kusuma KN.
KN Evidentiary value and
effects of contaminants on blood group factors in medico-legal
medico
grounds.. IAIM, 2015;
201 2(3): 68-72.

Available online at www.iaimjournal.com

Received on: 17-02-2015

Accepted on: 27-02-2015

Abstract
Background: Blood is considered to be the most common trace evidence obtained from the scene of
crime. Blood grouping reveals vital information regarding identification, in relation to crime
investigation. With this respect contamination
contamination with dust, rust, bacteria and detergent will have any
effect on blood grouping was studied.
Material and methods:: Randomly A, B, AB, Rh positive samples were taken in duplicates and were
added with rusted pins, dust particles, detergent, staphylocopcci bacteria and without any addition
one tube was kept in room temperature. Controls were kept in refrigerator at 44-8 C to compare the
results.
Results: There were lots of changes in blood groups after 10 days of incubation in all tubes with
contamination. Blood samples in dust after 10 days incubation had changed from A, B, AB to O blood
group with Rh negativity. Blood samples in rust had retained with original ABO group but Rh
positives were converted to Rh negative. Detergent tubes had also changed Rh positives
pos
to negatives
retaining original ABO. There was no change in either ABO or Rh blood groups in tubes with
Staphylococci bacteria.
Conclusion: Stains contaminated with common agents like dust, rust, bacteria and detergent give
inconclusive results by mixed
ixed agglutination method with passage of time. The results may be
accepted with caution, as the opinion might be medico legally significant.

International Archives of Integrated Medicine, Vol. 2, Issue 3, March, 2015.

Copy right 2015,, IAIM, All Rights Reserved.

Page 68

Effects of contaminants on blood group factors

ISSN: 2394-0026 (P)

ISSN: 2394-0034 (O)

Key words
Blood group, Contamination, Medico-legal.
Medico

Introduction
Human identification is based on blood type
which is determined by the presence or absence
of certain identifiers on the surface of red blood
cells. These identifiers, also
o called antigens,
identifies own red blood cell type. There are
four main ABO blood type groupings: A, B, AB,
and O.. These blood groups are determined by
the antigen on the blood cell surface and the
antibodies present in the blood plasma. In
addition to the ABO group antigens, there is
another blood group antigen located on red
blood cell surfaces known as the Rhesus factor
or Rh factor, this antigen may be present or
absent from red blood cells [1].. The use of blood
groups and protein polymorphisms to identify
possible sources of human stains left at the
scene of a crime is still being practiced by
judiciary.
y. Blood grouping is still vital in
identifying an individual though newer DNA
typing methods are available in many scenarios
[2].
Blood is considered to be the most common
trace evidence obtained from the scene of
crime. Collection of bloodstain from the scene of
crime as well as from the victim and suspects is
an important procedure for crime investigation
[3]. Grouping of forensic blood stains is known
to be trustworthy, but for investigation of
transfusion reactions at necropsy whole red cells
must be used [4]. Grouping blood has no reliable
results if it is contaminated with various agents
like dust, rust, detergents or any bacterial
contamination. Also stored blood contaminated
may also yield erroneous results with passage of
time. These results will be definitely of value in
medico legal cases [3].

With this background, this study was taken up,

to study the effects of various contaminants on
blood stored and the blood grouping results.

Material and methods

This study was done in the Department of
Pathology, Mandya Institute of Medical
Sciences, Mandya. Blood samples (5 ml) of
different blood groups were randomly collected
from blood bank attached to our college. A, B,
AB, O and Rh positives samples were taken
take in
sterile vacutainers in 6 duplicates.
One tube was kept as control which was stored
in refrigerator at 4-8 C. Second tube was kept at
room temperature. Third tube of all blood
samples were added with a pinch of dust. Fourth
tube was added with rusted pins of iron, fifth
tube with chunks of detergent and last tube of
all blood groups were added with
Staphylococcus species bacteria.
All the tubes were kept at room temperature
except the controls for 20 days. On the first day,
grouping was done and results
resu were noted. At
th
regular intervals on 5 day, 10th day, 15th day
and 20th day, blood grouping was done and
results were noted.
Blood grouping was done using slide
agglutination method with anti-A,
anti
anti-B, Rh
antisera using standard methods [1].

Results
Slide agglutination results were noted and
analyzed. On 1st day of blood grouping the
results were same as the controls without any
change in the blood groups. On 5th day, there
was no change in blood groups except for

International Archives of Integrated Medicine, Vol. 2, Issue 3, March, 2015.

Copy right 2015,, IAIM, All Rights Reserved.

Page 69

Effects of contaminants on blood group factors

weaker reactions in few blood samples
samp
from
tube with dust particles.
There were lots of changes in blood groups after
10 days of incubation in all tubes with
contamination which has been predicted as per
Table - 1.
Blood samples in dust after 10 days incubation
had given varied results, all blood groups (A, B,
AB) had changed to O blood group with Rh
negativity. Few blood samples in rust had
retained with original ABO group but Rh
positives were converted to Rh negative.
Detergent tubes had also changed Rh positives
to negatives retainingg original ABO. There was
no change in either ABO or Rh blood groups in
tubes with Staphylococci bacteria.

ISSN: 2394-0026 (P)

ISSN: 2394-0034 (O)
A, B, AB and Rh in blood contaminated with
dust, detergent and bacteria [3]. Our study
supported the findings of other authors who
strongly advised against the use of blood
grouping of decomposed and contaminated
stains, as the results may be inconclusive [5].
In contrast to one study [3], our study indicated
that bacterial contamination has no change in
any blood groups. This can be justified saying
that our blood is usually having transient
bacteremia with oral and skin colonisers
c
like
Staphylococci species which usually doesnt alter
the blood groups. A study reported the
existence of gram negative coccobacilli which
affected the blood group A substance and other
study have reported the effects of aerobic soil
microorganisms and cell free extracts, which
decomposes blood group substances [6, 7, 8].

Discussion
The reliability of blood grouping from
contaminated stains has been always
questioned in medico-legal
legal scenarios.
The tissues of the bodyy decompose at different
rates after death, but little is known of how long
red cell antigens persists [4]. Blood grouping is a
very important issue in crime scenes. Before
days only blood group identification was useful
method in fixing up the identity of criminal.
Though nowadays, DNA typing methods are
available, still blood grouping is important in
issues where there are any mismatched blood
transfusions and complications.
In this study, there was gross change in the
blood group after 10 days of blood storage
especially when contaminated with dust. There
was loss of A and B antigens giving the blood
group as O. Rh positive blood had turned up into
Rh negative in blood with dust, detergent and
rust after 10 days of storage. Another similar
study has shown
n similar report with changes in

The reports of false agglutinogens in stored

blood have appeared [4]. It becomes very
important that blood stains on rusted weapons
will just change in Rh factor. This has been
be in
contrast with a study which has said that rust
did not show any effect on the RBC or the
agglutinogenic capacity and grouping could be
satisfactorily done up to the third week [3]. Rh
factor appears more vulnerable to rust (ferric
oxide).
In detergent sample, there was change in Rh
factor after 10 days of incubation in B blood
group. Similar findings were noted in the sample
marked detergent the Rh was negative at 10th
day.
It
has
been
established
that
hemagglutination is dependent on the
membrane
rane flexibility and deformability of the
erythrocytes, which in turn is influenced by the
ATP content [9, 10]. Detergents contain alkaline
agents, which alter the fragility of the
erythrocyte membrane that in turn affects the
antigenicity [7].

International Archives of Integrated Medicine, Vol. 2, Issue 3, March, 2015.

Copy right 2015,, IAIM, All Rights Reserved.

Page 70

Effects of contaminants on blood group factors

Conclusion
Blood grouping though is an important identity
in identifying crime; it is not reliable if it is
contaminated with dust, detergent, rust etc.
Older stains in crime will be usually
contaminated. So we have to be careful while
identifying blood groups in case of medico-legal
medico
issues.

7.

8.

References
1. Geoff Daniels. ABO, H, and Lewis
systems. Human Blood Groups. 3rd
edition, Wiley-Blackwell,
Blackwell, 2013, p. 1-11.
1
2. Court D. S. Blood Grouping. Wiley
Encyclopedia of Forensic Science, 2009.
3. Chakraborty PC, Chattopadhyay S.
Effects
ffects of Common Contaminants on
Blood Group Factors in Medico-Legal
Medico
Ground. Indian Journal of Medical
Toxicology, 2011; 5(1): 73-74.
73
4. Enticknap JB. Survival of Blood Group
Factors After Death. Journal of Clinic
Path., 1957; 10: 199.
5. Getller AO, Kramer HE. Blood Grouping
in Forensic Medicine. The Journal of
Immunology, 1936; 31: 321-329.
321
6. Chase
MW.
A
Microorganism

Source of support: Nil

9.

10.

ISSN: 2394-0026 (P)

ISSN: 2394-0034 (O)
decomposing
group
specific
A
substances. J Bacteriol., 1938; 36: 383383
390.
Gilmore TE, Howe C. An Aerobic Soil
Microorganism which Decomposes
Blood
d Group Substances.
Substances I. Metabolic
and Immunochemical Studies. J
Bacteriol., 1959; 78(6): 806-813
806
.
Gilmore TE, Howe C. An Aerobic Soil
Microorganism which Decomposes
Blood Group Substances.
Substances II. Effects of
Cell free Extracts on Blood Group
Substances. J Bacteriol., 1959; 78(6):
814-820.
Tsuji T, Kimura A, Nishi K, Ito N,
Mizumoto J, Wada K. Effects of red cell
shape changes on haemagglutination.
Jpn J Legal Med., 1985; 39: 138-142.
138
Nishi K, Yamada M, Wakasugi C. Effects
of treatment with glutaraldehyde and of
storage in Acid- Citrate- Dextrose
solution
on
agglutinability
of
erythrocytes. Jpn J Legal Med., 1983; 37:
757-763.

Conflict of interest: None declared.

International Archives of Integrated Medicine, Vol. 2, Issue 3, March, 2015.

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Page 71

Effects of contaminants on blood group factors

ISSN: 2394-0026 (P)

ISSN: 2394-0034 (O)

Table 1:: Results of blood grouping in various tubes with different conditions.
Blood
group

AB

Days
1
5
10
15
20
1
5
10
15
20
1
5
10
15
20
1
5
10
15
20

Control
tube
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE

Room
temp
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE

Dust

Rust

Detergent

Staphylococci

+VE
+VE
O +VE
O VE
O VE
+VE
+VE
O VE
O VE
O VE
+VE
+VE
O VE
O VE
O VE
+VE
+VE
O VE
O VE
O VE

+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
B-VE
B-VE
+VE
+VE
AB-VE
AB-VE
AB-VE
+VE
+VE
+VE
+VE
+VE

+VE
+VE
+VE
+VE
+VE
+VE
+VE
B-VE
B-VE
B-VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE

+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE
+VE

International Archives of Integrated Medicine, Vol. 2, Issue 3, March, 2015.

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Page 72